Fig. 2.

Genetic variations in genomic DNAs from APO-SUS (S) and APO-UNSUS (U) (hypo)thalamus. Rats were from the original (F32) or the replicate (F18) rat lines. (A) Products generated by AP-PCR using primers AP-1 (product 1), AP-7 (product 2) or AP-777 (product 3) on genomic DNAs digested with the methylation-insensitive enzyme RsaI and the methylation-sensitive enzyme HpaII. ND = not determined. (B) Products 1, 2 and 3 generated by AP-PCR on genomic DNAs digested with RsaI and HpaII (H) or with the two methylation-insensitive enzymes RsaI and MspI (M). MspI products served as controls to determine whether products 1, 2 and 3 were due to differential methylation or to a genetic polymorphism. The fact that products 1, 2 and 3 were still present following MspI digestion indicates that they represent products without an HpaII site (“genetic fragments”). (C) Products generated by AP-PCR using a topoisomerase II binding site consensus sequence (5′-GCCTCCTTGCAGGTCTTT-3′) on genomic DNAs digested with the methylation-insensitive enzymes EcoRI (product 4) or MboI (product 5). Arrows indicate increased amounts of the AP-PCR products